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1.
为探讨不同有机溶剂对常用中草药的提取效果及其提取物对水产常见病原菌的抑菌活性，以50%乙醇、无水乙醇、氯仿、甲醇、丙酮、乙酸乙酯和石油醚为提取溶剂，根据水浴旋转蒸发器回流提取工作原理，分别对五倍子、乌梅、诃子、大黄和五味子等5种中草药进行固液萃取，并用其提取物对水产高致病性病原菌嗜水气单胞菌（Aeromonas hydrophila）进行体外抑杀菌试验。结果表明：50%乙醇、无水乙醇、甲醇对5种中草药的提取效果优于其他4种溶剂，尤以甲醇的提取效果最佳，其提取率分别为五倍子46.09%、乌梅27.58%、诃子29.55%、大黄50.34%、五味子30.88%;提取效果最差的是石油醚，其提取率分别为五倍子1.98%、乌梅1.73%、诃子2.21%、大黄0.89%、五味子3.40%。体外抑菌试验结果显示，嗜水气单胞菌对五倍子的3种醇提物均表现为极度敏感，对大黄的甲醇提取物表现为高度敏感，对五倍子、大黄和五味子的氯仿提取物以及对乌梅、大黄和五味子的石油醚提取物呈低度敏感，对五倍子的丙酮提取物等共25种提取物表现为中度敏感；五倍子、大黄的醇提物对供试菌的抑杀菌效果明显优于其他3种中草药的任何一种... 相似文献
2.
叶绿素a是水环境评价和水生态研究中的一项重要指标。针对叶绿素a测定中前处理耗时长、提取不完全等问题,研究了液氮冻融前处理对叶绿素a测定效率的影响。以实验室培养的小球藻(Chlorella pyrenoidosa)和微囊藻(Microcystis aeruginosa)藻液为实验材料,通过单因素实验和正交实验设计,探究速冻时间、冻融次数、浸提时间等主要参数对提取效率的影响,并确定液氮冻融前处理测定叶绿素a的最优条件。结果表明,液氮冻融前处理测定叶绿素a的最优条件为速冻30 s、冻融4次、90%的丙酮浸提4 h,在此条件下,小球藻和微囊藻叶绿素a浓度测定相对标准偏差分别为0.57%和2.25%,变异系数CV分别为0.46%和1.84%。野外水样测定结果显示,与丙酮-研磨法、分光光度法(SL 88-2012)、分光光度法(HJ 897-2017)和反复冻融浸提法相比,液氮冻融前处理法的提取效率高于其他方法且测定结果相对偏差更小,浸提时间较其他4种方法缩短50%以上,其浸提效率达93%以上。液氮冻融前处理法对叶绿素a的提取效率明显高于其他方法且数据重现性较好,具有操作简单、结果准确、操作耗时短等特点。 相似文献
3.
鲢肌肉脂肪酸分离方法效果比较及组成特征分析 总被引：1，自引：0，他引：1
采用了三种不同方法,即超声波提取法、索氏提取法和氯仿-甲醇法,从鲢(Hypophthalmichys molitrix)肌肉中提取鱼油,继而进行酸性或碱性甲酯化,经气相色谱/质谱联用技术,对其脂肪酸组成特征进行了定性和定量分析。结果显示:鲢肌肉检测出25种脂肪酸,其中包括7种饱和脂肪酸,8种单不饱和脂肪酸和10种多不饱和脂肪酸。获得不饱和脂肪酸含量占总脂肪酸含量分别为41.39%、36.15%、33.55%,可见三种分离方法间存在明显分离效果差异。因此得出石油醚作溶剂的超声波萃取法是提取和分析鱼类脂肪酸较理想的方法。 相似文献
4.
Consuelo Cháfer‐Pericás Ángel Maquieira Rosa Puchades Begoña Company Javier Miralles Amelia Moreno 《Aquaculture Research》2010,41(9):e217-e225
An analytical method based on HPLC with MS/MS detection was developed and optimized in order to determine the most useful antibiotics (sulphonamides and tetracyclines) used in aquaculture. A simple extraction procedure, without any clean‐up step, was evaluated in order to obtain maximum analyte recovery from fish samples (Sparus aurata). A mixture of methanol:water 70:30 (v/v)+1 mL EDTA 0.1 M was selected as optimum extractant solution. Because no matrix effects were observed, a standard calibration curve prepared in mobile phase was used for quantification purposes. Antibiotic‐free fish samples were spiked at different concentration levels and analysed by the optimized HPLC method. The average recoveries (n=6) obtained were satisfactory, ranging from 88% to 110% at 100 μg kg?1. The proposed methodology provided limits of detection for the tested antibiotics in the 1.2–16 μg kg?1 range, lower than 100 μg kg?1, the maximum residue level established by the European Union. Finally, commercial fish samples from different origins were analysed in order to confirm the usefulness of the developed methodology. 相似文献
5.
为研究辽东湾海域水体中叶绿素a含量的分布特征,于2013年8月(夏季)和12月(冬季)在辽东湾设置38个站位,分别采集了表、中、底层的海水样品,检测夏季和冬季两个航次水体中叶绿素a的含量,分析该海域叶绿素a的季节分布与空间变化特征。同时利用相关性统计分析的方法,将各水质因子对叶绿素a含量影响的差异显著性进行了分析。结果显示,调查海域夏季的叶绿素a平均质量浓度为6.90μg/L,冬季为1.26μg/L,时间分布上夏季质量浓度明显高于冬季;空间水平分布上,夏季辽东湾湾口的平均叶绿素a质量浓度明显高于湾内的平均质量浓度,而冬季则越靠近湾口处叶绿素a的平均质量浓度越低,与夏季规律正好相反;空间垂直分布上,夏季表层叶绿素a的质量浓度最高,10m层次之,底层的质量浓度最低;而冬季的叶绿素a质量浓度则呈现10m层最高,底层次之,表层质量浓度最低的趋势。说明辽东湾叶绿素a含量存在明显的时间和空间分布特征;结合同期的温度、盐度、溶解氧等环境因子的调查结果,统计分析结果表明调查海域中环境因子温度、盐度、溶解氧和营养盐的季节性变化是导致叶绿素a含量趋势变化的最可能原因之一。 相似文献
6.
A Strategy for Sperm Cryopreservation of Atlantic Salmon,Salmo salar,for Remote Commercial‐scale High‐throughput Processing 
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下载免费PDF全文 Huiping Yang E Hu John T. Buchanan Terrence R. Tiersch 《Journal of the World Aquaculture Society》2018,49(1):96-112
Sperm cryopreservation is an essential tool for long‐term storage of genetic resources for aquaculture fishes. The goal of this study was to develop an efficient and streamlined protocol for high‐throughput processing for sperm cryopreservation in Atlantic salmon, Salmo salar. The objectives were to evaluate: (1) osmolality of blood serum for determining extender osmolality, (2) effects of extenders for fresh sperm dilution and refrigerated storage, (3) effects of methanol and dimethyl sulfoxide (DMSO) on fresh sperm motility, and (4) motility and fertility after thawing. In this study, sperm samples were collected at a hatchery site in Canada and shipped to a freezing site located 2200 miles (3550 km) away in the USA. Evaluation of three extenders indicated that Mounib solution was suitable for diluting dry sperm for sample processing. Ten percent of methanol or DMSO was less toxic to sperm cells than was 15% within 30 min. Further testing with methanol at 5, 10, and 15%, and sperm solution : extender dilutions (v:v) of 1:1, 1:3, and 1:19 (at concentrations of 5 × 107, 3 × 108, and 1 × 109 cells/mL) indicated that methanol at 5 and 10% showed less toxicity to fresh sperm within 1 h at sperm:extender dilutions of 1:1 and 1:3. Post‐thaw motility of sperm cryopreserved with 10% methanol was significantly higher than that with 10% DMSO, and fertility reflected those results (0–1% in DMSO vs. 38–55% in methanol). Further evaluation of sperm cryopreservation with 10 and 15% methanol at sperm dilution ratios of 1:1, 1:3, and 1:19 indicated that post‐thaw motility in 10% methanol was significantly higher than that in 15% methanol, and post‐thaw fertility in 10% methanol at 1:1 and 1:3 dilution ratios had fertilization rates similar to that of fresh sperm controls. Sperm samples from 12 males cryopreserved with 10% methanol showed male‐to‐male variation in post‐thaw motility (0–36%). Overall, a simplified standard protocol was established for cryopreservation of shipped sperm of Atlantic salmon using extender without egg yolk and yielded satisfactory post‐thaw motility and fertilization rates. This procedure can be readily adopted by aquaculture facilities to take advantage of high‐throughput cryopreservation capabilities at remote service centers. Most importantly, this approach lays the groundwork for an alternative commercial model for commercial‐scale production, quality control, and development of industrial standards. Control of male variability and sperm quality remain important considerations for future work. 相似文献
7.
以驼海燕为原料,用丙酮沉淀法制备蛋白酶粗酶,并对其酶学性质进行研究。试验结果表明,提取蛋白粗酶的最佳条件为:原酶液与丙酮的体积比为1∶1.6;蛋白酶反应的最适温度为45℃,最适pH值为8.1;pH为7～9时可保持较高的酶活力;蛋白酶的酶活力在20～50℃基本稳定。酶促反应结果表明,Na+、K+、Li+、Mg2+对蛋白酶活力具有促进作用,而Zn2+、Cu2+、Hg2+能抑制蛋白酶的活力,EDTA对蛋白酶活力的影响甚微。酶动力学方程表明,最大反应速度vmax=5.99U/mL,米氏常数Km=0.69g/mL。 相似文献
8.
针对2007-2009年间在黄海海域形成"绿潮".的大型绿藻浒苔(Enteromorpha prolifera).研究了其3种不同极性有机溶剂(甲醇、丙酮、氯仿)提取物对太平洋牡蛎(Crassostrea gigas)受精卵孵化的影响.结果表明.在48 h内.浒苔的甲醇和丙酮提取物对太平洋牡蛎受精卵孵化有较强的抑制作用.高浓度提取物能够完全抑制太平洋牡蛎受精卵孵化至D型幼虫.而氯仿提取物对太平洋牡蛎受精卵孵化的影响没有明显的浓度-效应关系.应用乙酸乙醋和石油醚对浒苔的甲醇提取物进行液-液萃取.并探讨了乙酸乙醋相和石油醚相中的浒苔提取物对牡蛎受精卵孵化的影响.结果表明.石油醚相和乙酸乙醋相中的浒苔提取物对牡蛎受精卵孵化均有很强的抑制效应.上述研究表明.浒苔中存在能够抑制太平洋牡tfi受精卵孵化的活性物质.这类物质应具有较高的极性. 相似文献
9.
运用加速溶剂萃取技术（accelerated solvent extraction,ASE）,通过单因素试验和响应面分析法优化海刀豆（Canavalia maritima）中总黄酮的提取工艺。在单因素试验的基础上,选取萃取温度、萃取时间和甲醇体积分数3个因素为自变量,总黄酮提取率为响应值,采用Box-Behnken试验设计方法,研究各因素的显著性及其交互作用对总黄酮提取率的影响。利用Design Expert软件,建立了提取率与各因素的二次多项式数学模型,得到了海刀豆中总黄酮的最佳提取条件：萃取压力10.3 MPa,萃取温度130 ℃,萃取时间16 min,甲醇体积分数66%,循环次数2次,总黄酮提取率为83.43%,与理论值相对偏差-1.63%。 相似文献
10.
坛紫菜叶状体蛋白质双向电泳样品制备方法的比较 总被引：1，自引：0，他引：1
蛋白质样品的制备是双向电泳分析的关键。为探索适用于坛紫菜叶状体蛋白质双向电泳样品的制备方法,实验对3种常用植物蛋白质分离方法进行了优化和改进,并分别同3种蛋白质裂解液联用,比较了9种方法的总蛋白质得率及双向电泳图谱效果。结果发现,采用三氯乙酸/丙酮沉淀法与裂解液C(7 mol/L 尿素,2 mol/L 硫脲,2%CHAPS,2%TritonX-100,2% IPG Buffer pH 3~10,65 mmol/L DTT)联用的方法,坛紫菜叶状体的蛋白质得率最高,且双向电泳图谱显示此法获得的蛋白质点数最多,蛋白点形状规则,水平和垂直拖尾情况较轻,图像清晰。实验结果还发现,在进行双向电泳分析时,采用pH 4~7的胶条可以使坛紫菜叶状体蛋白质得到更为有效的分离。 相似文献
11.
通过萃取回收和色谱条件优化,建立了运用高效液相色谱法(HPLC)检测鳗鲡血浆和肌肉中乙胺嘧啶的方法。结果显示:在柱温30℃、检测波长236 nm、流动相乙腈、水、乙酸体积比为15∶85∶0.25和流速1.0 mL/min的色谱条件下,在设置鳗鲡血浆及肌肉中乙胺嘧啶浓度分别在0～5.0μg/mL与0～2.0μg/g范围内,相关曲线方程分别为y=766.68x+9.2511与y=853.96x+8.0956,相关系数(r)分别为0.9998和0.9999。在萃取回收中,向血浆和肌肉样品中分别加入适量无水硫酸钠,再分别用乙腈、三氯甲烷体积比为10∶1和5∶5的混合液萃取两次,回收率均达80%以上,日间、日内变异系数均低于3%,最低检测限可分别达0.005μg/mL和0.005μg/g。 相似文献
12.
Li Zhou Peixuan Li Yuling Zhao Shuang Hou Baolei Cong Jun Huang 《Journal Of Aquatic Food Product Technology》2017,26(7):824-834
Response surface methodology was applied to determine optimal conditions for lipid extraction from shrimp. Box-Behnken Design including independent variables such as extraction time, extraction temperature, and ratio of mixed solvents was used. The optimal conditions for lipids extraction were the use of petroleum ether:ethyl acetate (2.3:1, v/v) with 62 min of extraction at 52ºC. Under the optimal conditions, the extraction yields of lipids from Euphausia superba, Penaeuschinesis, Macrobranchium nipponense, and M. rosenbergii were 4.39, 2.80, 2.98, and 3.30%, respectively. Gas chromatographic analysis showed that M. rosenbergii presented the highest level of polyunsaturated fatty acid (PUFA) (48.28%) among the shrimps studied. In addition, the 20:5n3 (EPA) + 22:6n3 (DHA) value (26.43%) was second only to E. superba (31.27%). The finding suggests that M. rosenbergii is a potential PUFA source for food supplementation. 相似文献
13.
目的:建立SPE-HPLC分析方法同时测定饲料中己烯雌酚、己烷雌酚、双烯雌酚和雌二醇。方法:饲料样品经甲醇水溶液提取,正己烷脱脂,HLB固相萃取小柱净化后用HPLC分离测定。采用Xterra RP18色谱柱,乙腈-0.025% H3PO4(48:52)为流动相,检测波长228nm。结果:己烯雌酚、己烷雌酚、双烯雌酚和雌二醇的检测限分别为0.02、0.02、0.01、0.02μg/mL;线性范围均为0.1～5.0μg/mL;萃取回收率≥80.5%;日间精密度≤2.5%。结论:所建方法准确、灵敏、简便,适用于饲料中4种雌激素的分析。 相似文献
14.
鱼肉和饲料中的虾青素分析方法对于研究鲑鱼肉色质量及饲料对其影响至关重要。本研究建立了北极红点鲑鱼肉类胡萝卜素的正相高效液相色谱分析方法。该方法采用正己烷与异丙醇等度洗脱(93∶7,体积比),流速1.1mL/min,可实现18min内4种常见鲑鱼类胡萝卜素包括虾青素、角黄素、β-胡萝卜素和玉米黄素的有效分离。研究结果显示,虾青素的回收率和变异系数值分别为97%和1.5%。正己烷和异丙醇复合溶剂提取的鱼肉样品与丙酮提取并经氨基固相萃取小柱纯化后的鱼肉样品的虾青素含量接近,但比丙酮提取样品中虾青素含量略低。正相高效液相色谱法和分光光度法测定鱼肉虾青素含量分别为1.43mg/L和2.43mg/L。研究同时建立了鲑鱼饲料虾青素分析方法。研究结果表明,-80℃冷冻结合氨基固相萃取小柱纯化可去除饲料样品中的大部分脂肪。饲料中虾青素酯皂化的最佳条件为:0.10mol/L甲基化氢氧化钠,22℃,20min;或者0.12mol/L甲基化氢氧化钠,4℃,3h。研究结果表明,两种方法可用于鲑鱼肉和饲料中的虾青素分析,为相关研究带来方便。 相似文献
15.
Antioxidant activity of the marine alga Kappaphycus alvarezii was investigated. Methanol, acetone, petroleum ether, aqueous methanol, ethyl acetate, and chloroform extracts (1 mg/mL) of K. alvarezii were tested for their 2, 2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity. The values were compared with those of Vitamin C and butylated hydroxytoluene (BHT). Extracts showing positive results, when tested for DPPH free radical scavenging, were examined for dose effect, in-vivo hydrogen peroxide (H2O2) and hydroxyl radical scavenging assays. All extracts showed dose-dependent DPPH scavenging and significant hydroxyl radical scavenging activities (> 82.6%). The acetone, aqueous methanol, and methanol extracts of K. alvarezii showed the highest scavenging activity. Ethyl acetate extract showed a moderate activity of 62.9%. In the DPPH method, petroleum ether and hexane extracts showed less activity with IC50 values of 118.58 ± 8.94 and 116.25 ± 7.14 μg/mL, respectively. Acetone, methanol, and ethyl acetate extracts exhibited IC50 values of 57.32 ± 1.07, 61.31 ± 0.67, and 79.50 ± 1.59 μg/mL, respectively. K. alvarezii showed higher antioxidant activity in both in vitro and in vivo studies. Proton NMR studies revealed signals in the region 0.5 to 2.0 ppm suggesting the presence of steroidal identity in the extracts. 相似文献
16.
Abstract. This paper reports a series of trials on the cryopreservation of Oreochromis niloticus (L.) spermatozoa, using methanol as the cryoprotectant.
Immotile milt samples pooled from four males were diluted with two diluents, each being subjected to equilibration periods of 15, 30, 45, 60 and 90 min. In addition, samples of fresh pooled milt were kept for 0, 2, 4, 6 and 8 days at 4°C prior to cryopreservation. Diluted samples were stored in 250-μl plastic straws and cooled to -50°C at 5°C/min and held under liquid nitrogen for between 1 and 3 weeks. Viability of post-thawed spermatozoa was estimated from video recordings of samples activated under a microscope and from fertilization rates of eggs.
The type of diluent and its interaction with equilibration time significantly (P< 0.05) influenced the post-thaw motility of spermatozoa. Spermatozoa stored for up to 6 days prior to cryopreservation were as fertile as freshly frozen spermatozoa and yielded 60.6 (±SE, 8.4) developing embryos. 相似文献
Immotile milt samples pooled from four males were diluted with two diluents, each being subjected to equilibration periods of 15, 30, 45, 60 and 90 min. In addition, samples of fresh pooled milt were kept for 0, 2, 4, 6 and 8 days at 4°C prior to cryopreservation. Diluted samples were stored in 250-μl plastic straws and cooled to -50°C at 5°C/min and held under liquid nitrogen for between 1 and 3 weeks. Viability of post-thawed spermatozoa was estimated from video recordings of samples activated under a microscope and from fertilization rates of eggs.
The type of diluent and its interaction with equilibration time significantly (P< 0.05) influenced the post-thaw motility of spermatozoa. Spermatozoa stored for up to 6 days prior to cryopreservation were as fertile as freshly frozen spermatozoa and yielded 60.6 (±SE, 8.4) developing embryos. 相似文献
17.
To develop a suitable cryopreservation diluent for spermatozoa of the endangered Sakhalin taimen Hucho perryi, all possible combinations of cryoprotectants (glycerol, dimethyl sulfoxide [DMSO], methanol) and extenders (fetal bovine
serum [FBS], 300 mM glucose solution [GS], artificial seminal plasma for masu salmon) were examined by observing sperm motility
10 s after thawing. Spermatozoa cryopreserved with diluents such as mixtures of 10% glycerol plus 90% FBS, 10% DMSO plus 90%
FBS, and 10% methanol plus 90% GS showed the highest motility. The maximal post-thaw motility was observed at 10% among all
concentrations (0, 5, 10, 15 and 20%) of these three cryoprotectants. No significant difference among three diluents was observed
in motility at 10 s. Mixtures of 10% glycerol plus 90% FBS, 10% DMSO plus 90% FBS, and 10% methanol plus 90% GS are suitable
cryopreservation diluents for Sakhalin taimen spermatozoa. 相似文献
18.
Maturation-inducing steroid (MIS) in the Indian female catfish,Clarias batrachus, was purified and characterized from the incubation medium in which fully grown but immature folliculated oocytes were incubated with salmon gonadotropin (SG-G100) for 36 h. Maturation-inducing (MI) activity of residues obtained at various steps of extraction and purification was assessed byin vitro germinal vesicle breakdown (GVBD) assay using folliculated oocytes ofC. batrachus. The post incubation medium was extracted with diethyl ether. The ether phase was partitioned using 50% methanol plus n-hexane. The methanol phase which had MI activity was fractionated into 7 fractions using reverse-phase high-performance liquid chromatography. Of these 7 fractions, fraction 3 was found to be active in having MI ability and identified as 17 ,20-dihydroxy-4-pregnen-3-one (17,20-diOHprog). The authenticity of 17,20-diOHprog as the major follicular mediator of gonadotropin-induced oocyte maturation was further confirmed by thin-layer chromatography (TLC) in which fraction 3 was run along with authentic 17,20-diOHprog standard. This investigation gives a direct evidence that 17,20-diOHprog is the major naturally occurring MIS in Indian female catfish,C. batrachus. 相似文献
19.
建立了渔业环境沉积物中硫丹及其代谢物硫丹硫酸酯残留量的气相色谱分析方法。样品以正已烷/丙酮(1∶1,V/V)提取、铜粉除去硫化物、石墨化碳黑-佛罗里硅土复合固相萃取柱净化、二氯甲烷/正已烷(3∶7,V/V)洗脱,用电子捕获-气相色谱(GC-ECD)测定,外标法定量。硫丹各组分在0.001～0.2μg/mL浓度范围内线性相关系良好(r≥0.9989);1、10、20μg/kg添加浓度的回收率在76.3%～107%之间,相对标准偏差(RSD)为2.31%～11.8%;α-硫丹、β-硫丹、硫丹硫酸酯的检出限(LOD)分别为0.03、0.04、0.05μg/kg(干重),最低定量限(LOQ)均为1μg/kg(干重)。适用于渔业环境硫丹污染的监测。 相似文献
20.
为了有效提取营养功效和附加值均更高的磷虾油,对南极磷虾(Euphausia superba)资源的综合开发利用提供参考,利用2013年7-8月和2013年12月-2014年5月在南乔治亚群岛和南设得兰群岛附近海域采集的南极磷虾样品,采用索氏抽提法,分别用无水乙醇、丙酮、石油醚、正己烷、乙酸乙酯、环己烷6种常见的有机溶剂作为提取试剂,对南极磷虾体内的脂肪进行提取,并确定最佳萃取溶剂.同时对南极磷虾脂肪、蛋白含量的季节变化及其与体长的关系进行了分析.结果显示,无水乙醇为南极磷虾脂肪提取的最佳溶剂.南乔治亚群岛海域7-8月南极磷虾脂肪含量和蛋白含量(干重)分别为(22.03±0.51)％和(64.52±0.16)％;其中,全长小于40 mm的磷虾个体,脂肪与蛋白含量均显著高于全长大于40 mm的个体(P＜0.05).从南极的初夏到秋末,南设得兰群岛海域的南极磷虾脂肪含量呈逐渐上升趋势,由12月的(19.83±0.04)％上升至翌年4月的最大值(30.584±0.02)％;5月则略有降低,为(28.66±0.03)％.与此相反,南极磷虾的蛋白含量则是12月最高、4月最低,分别为(66.75±0.14)％和(58.02±0.23)％;脂肪含量与蛋白含量呈显著负相关(P=0.005).研究结果对南极磷虾渔业及其生产安排具有直接的指导意义. 相似文献